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14FARMACJA | ФАРМАЦЕВТИЧЕСКИЕ НАУКИ
DEVELOPMENT AND VALIDATION OF METHODS OF ASSAY OF LORATADINE IN
TABLET
O.B. Polyauk
PhD, Head of Pharmaceutical Chemistry Department Ternopil State Medical University named after I. Ya. Horbachevsky (Ternopil, Ukraine)
D.B. Korobko
PhD, Dean of Pharmaceutical faculty, associate professor of Pharmaceutical Chemistry Department Ternopil State Medical University named after I. Ya. Horbachevsky (Ternopil, Ukraine)
L.S Logoyda
PhD, Vice Dean of Pharmaceutical faculty, associate professor of Pharmaceutical Chemistry Department Ternopil State Medical University named after I. Ya. Horbachevsky (Ternopil, Ukraine)
N.O.Zarivna
PhD, associate professor of Pharmaceutical Chemistry Department Ternopil State Medical University named after I. Ya. Horbachevsky (Ternopil, Ukraine)
ABSTRACT
The current pharmaceutical analysis has got more emphasis to satisfy our query for better understanding of physico-chemical properties of pharmaceutical compounds, by the use of advanced instrumental methods. The pharmaceutical industry is under increased scrutiny to constrain costs and yet consistently deliver to market safe, efficacious products that fulfill medical needs. As a part of this, drug analysis also plays an important role. Standard analytical procedure for newer drugs or formation may not be available in Pharmacopoeia; it is essential to develop new analytical methods which are accurate, precise, specific, linear, simple and rapid. Development and testing of new analytical methods represents our primary aim. Method of assay of loratadine in tablets by absorption spectrophotometry has been developed and validated. The basic validation parameters were specificity, linearity, range, accuracy (convergence), accuracy and robustness. Since fulfilled the eligibility criteria in respect of performance validation in accordance with the requirements of SPU, the method can be used for the determination of loratadine in tablets and can be included in the quality control methods for loratadine in tablets.
Keywords: loratadine, active pharmaceutical ingredient (API), tablets, spectrophotometry, validation.
Loratadine - one of the few medicines that can solve the besides API, various excipients differently will influence the problem of allergies complex. Worldwide, it has established itself analysis. Also analyzed the literature data, quantification of as the most effective and safest antihistamine agent. Together loratadine in medicines by spectrophotometric methods is with the Hl-blocking effect it has many additional anti-allergic almost not described. Therefore, the above mentioned methods properties that are important in clinical practice because it has we have chosen to further study the quantitative determination a wider range of therapeutic possibilities in the treatment of of loratadine in tablets. The problem of the development of various allergic diseases, to avoid the appointment of a large new methods for the quantitative determination of active number of other drugs. It's used to quickly achieve remission pharmaceutical ingredients in medicines was and is still a and relapse prevention spending allergic diseases, especially in problem today. Therefore, the development of new, more children [1] and in treatment of complex allergic diseases such advanced and simple in execution methods of analysis of as asthma and allergic rhinitis, allergic conjunctivitis, atopic medicines remains a problem of modern pharmacy. dermatitis and others. The purpose of the work was development and validation
In the pharmaceutical market of Ukraine presented as of spectrophotometric method of quantitative determination mono-drugs or some combination medicines containing of loratadine in tablets. loratadine. Experimental
Substance loratadine is among SPU monographs. Materials and methods According to SPU identification is carried out by absorption The study was conducted on five samples of domestic spectrophotometry in the infrared region of the spectrum [2]. producers loratadine tablets: «Zdorovja» (series 31012), Quantitative determination of loratadine in substance carried «Lekchim» (series 100512), «Farmak» (series 330912), out by potentiometric titration. Due to the use oflarge quantities «Arterium» (series 40612), «Darnitsa» (series 200712). This of organic solvents for the test, complexity and sustainability used analytical equipment: spectrophotometer Carry 50, sample preparation and methods in general, the high cost of electronic scales AVT-120-5DM; measuring vessel class, reagents and instruments - will increase the costs of analysis appropriate reagents that meet the requirements of SPU and create difficulties in the analysis, as the SFS contains and pharmacopoeia standard sample (CRS) of loratadine
(certificate number 11 / 1-243 of 02.12.13), acquired in the State Enterprise «Ukrainian Scientific Center pharmacopoeia quality of medicines».
Results and Discussion
Preliminary tests we have received ethanol and hydrochloric extracts of the research object that spectrophotometried and got absorption maxima. UV spectrum of ethanol extract of loratadine tablets has expressed intense absorption band at 248 ± 2 nm and hydrochloric removal of loratadine tablets -248 nm ± 2 and 274 ± 2 nm, respectively [1]. As a result of the experiment, we selected the proper conditions of sample preparation for quantitative determination of loratadine in tablets that let you choose the optimum ethanol as extragents because through him we got a pronounced absorption maximum in comparison with hydrochloric withdrawal.
Methods of quantitative determination of loratadine tablets.
Tested solution. The exact weight powdered tablets, equivalent to 0.0125 g of loratadine, placed in a volumetric flask 50.0 ml, add 20 ml of 96% alcohol R, dissolved within 5-10 minutes when heated in a water bath at 50 ° C, cooled and adjusted volume solution of 96% alcohol R to tags, stirred and filtered. 1.0 ml of this solution proves 96% alcohol R to 25.0 ml.
Reference solution. 12.5 mg loratadine CRS dissolved in 50.0 ml of 96% alcohol R. 1.0 ml of the solution adjusted 96%
alcohol R to 25.0 ml.
Compensation solution. 96% alcohol R.
Absorbance of the test solution and reference solution is measured at a wavelength of 248 nm regarding compensation solution.
Calculate the content of loratadine in one tablet, in g, based on the average tablet weight, based on the content specified in CRS loratadine.
Content loratadine tablets, g, calculated using the formula:
X
A • m • m„
x o
A • m. t
o tot
where
A - optical density of the reference solution; A - optical density of the tested solution; m - weight of CRS loratadine sample, g; mtot- weight of powder sample pounded tablets, g; mavr - average weight tablets, g.
Content of loratadine should be 0,009 - 0,011 g in terms of the average weight of one tablet [3]. The results of the determination the content of loratadine in tablets «Loratadine» («Arterium») are shown in table 1.
Table 1
The results of the determination the content of loratadine in tablets «Loratadine» («Arterium»)
Mass of sample, g The optical density Ai Loratadine concentration in solution, mg / cm3 Found loratadine in one tablet, mg Metrological characteristics (P = 0.95 n = 5)
0,125 0,576 0,100 9,989 m = 9,983mg S=0,048 t=2.78 Am=0.6mg m=9.983±0.06mg £=0.6%
0,125 0,575 0,100 10,000
0,126 0,577 0,101 9,937
0,125 0,576 0,100 9,937
0,124 0,575 0,099 10,052
The results of the trial indicate that none of the results of We also analyzed loratadine in tablets and other domestic the largest error has not gone declared content of 0,009 - 0,011 manufacturers such as «Zdorovja», «Lekchim», «Farmak», g (table 1). «Darnitsa». The results of the tests are presented in table 2-5.
Table 2
The results of the determination the content of loratadine in tablets «Loratadine» («Zdorovja»)
Mass of sample, g The optical density Ai Loratadine concentration in solution, mg / cm3 Found loratadine in one tablet, mg Metrological characteristics (P = 0.95 n = 5)
0,150 0,518 0,120 8,993 m = 9,19mg S=0,399 t=2.78 Am=0,49 mg m=9,19±0,49mg £=5.33%
0,151 0,519 0,121 8,950
0,150 0,519 0,120 9,901
0,149 0,518 0,119 9,053
0,149 0,518 0,119 9,053
Table 3
The results of the determination the content of loratadine in tablets «Loratadine» («Farmak»)
Mass of sample, g The optical density Ai Loratadine concentration in solution, mg / cm3 Found loratadine in one tablet, mg Metrological characteristics (P = 0.95 n = 5)
0,240 0,557 0,192 9,670 m = 9,673 mg S=0,0411=2.78 Am=0,052mg m=9,673±0,052mg £=0.54%
0,241 0,558 0,193 9,647
0,238 0,556 0,191 9,734
0,240 0,558 0,192 9,687
0,242 0,559 0,194 9,625
Table 4 The results of the determination the content of loratadine in tablets «Loratadine» («Lekchim»)
Mass of sample, g The optical density Ai Loratadine concentration in solution, mg / cm3 Found loratadine in one tablet, mg Metrological characteristics (P = 0.95 n = 5)
0,170 0,530 0,136 9,201 m = 9,215mg S=0,047 t=2.78 Am=0,059mg m=9,215±0,059mg £=0.64%
0,171 0,531 0,137 9,165
0,171 0,532 0,137 9,182
0,168 0,528 0,134 9,276
0,169 0,530 0,135 9,255
Table 5 The results of the determination the content of loratadine in tablets «Loratadine» («Darnitsa»)
Mass of sample, g The optical density Ai Loratadine concentration in solution, mg / cm3 Found loratadine in one tablet, mg Metrological characteristics (P = 0.95 n = 5)
0,150 0,535 0,120 9,288 m = 9,317mg S=0,058 t=2.78 Am=0,073mg m=9,317±0,058mg £=0.62%
0,150 0,536 0,120 9,305
0,148 0,534 0,118 9,396
0,149 0,535 0,119 9,351
0,151 0,536 0,121 9,244
Every developed method of quantitative determination must be validated in accordance with the requirements of SPU. We conducted validation of methods of quantitative determination of loratadine in tablets.
Validation of analytical methods - is experimental evidence that the method is suitable for the task [3]. The validation was performed on the main validation parameters: specificity, linearity, precision, accuracy, robustness, range of application.
Linearity, convergence, accuracy and range of application methods have been studied in model solutions loratadine [3-6].
Research of the specificity When photometric solution obtained with ethanol extraction model tablet mixture containing adjuvants and
prepared similar test solution, absorption was observed at wavelength (248 ± 2 nm), i.e placebo tablets analyzed in terms of quantitative determination prevents absorption bands, and thus does not affect the optical density in the quantitative determination.
Research linearity methods
Linearity studied within 50-150% of nominal concentrations of loratadine. Solutions with exactly known concentration obtained by diluting the appropriate standard solution and perform general procedure for the determination. Based on the data, building plot of optical density on the concentration of loratadine in normalized coordinates and calculated linear parameters (table 6).
Table 6
The results of the studying of linearity of quantification of loratadine in tablets
№ Ci, mg / ml Cnorm, % A Anorm Requirements and eligibility criterion
1 3,75 50 0,228 49,9 equation of the line: Anorm = 0,078^Cnorm - 0,156 Slope: b = 0,078 The point of intersection with the axis of ordinates: a = -0,156 The correlation coefficient: r = 0,9999
2 4,50 60 0,275 60,2
3 5,63 75 0,340 74,4
4 6,75 90 0,411 89,9
5 7,50 100 0,457 100,0
6 8,63 115 0,526 115,1
7 9,38 125 0,572 125,2
8 10,50 140 0,637 139,4
9 11,25 150 0,687 150,3
The correlation coefficient: r = 0,9999 r < 0,99974 performed
The residual variance = 0,1329 Residual standard deviation = 0,3903 S0/b < 1,267 performed
The confidence interval constant a: Aa = 0,9229 a = - 0,1773 Aa < 1,536 performed statistical criteria and practical significance
Calculation of the linear relationship (according to the table 6) to loratadine was conducted by the least squares method, according to the requirements of SPU [4]. As a result of studies, we found that the method characterized strict linearity, correlation coefficient greater than 0.99974 (criterion value), a small residual standard deviation over a wide range (from 50 to 150% in normalized coordinates). Graph of the optical density of the solution concentration is almost through the origin (the point of intersection with the axis of ordinates -0.156), indicating no effect of background or excipients for optical density test solution in these conditions, and calculated confidence interval constant Aa confirms statistical and practical constant and insignificance.
Research precision
Precision expresses the degree of proximity or degree
of spread of results for a series of measurements made by this method in different samples of the same homogeneous sample. Table 7 shows the results of the research accuracy and convergence techniques. The results point to a convergence of methodology, its reasonable accuracy, because none of the results has an error more than 4.8% (complete uncertainty analysis result AAs = 4,8 with deviations content in compliance with SPU).
Research correctness
The tests were performed on standard solutions loratadine by «input-output». The test results are shown in table 7. As follows from the resulted results performed insignificance criterion practical bias, indicating that the latter is insignificant in comparison with a maximum uncertainty analysis.
Table 7
Results of testing methods for the quantitative determination of loratadine accuracy and convergence
№ Ci input, % Ci output, % Z=Ci output •Ш / Ci input , %
1 50 49,9 99,8
2 60 60,2 100,3
3 75 74,4 99,2
4 90 89,9 99,9
5 100 100,0 100,0
6 115 115,1 100,1
7 125 125,2 100,2
8 140 1 39,4 99,6
9 150 150,3 100,2
Arithmetic mean, Z 99,9
The relative standard deviation, Sz% 0,35
Continue
The relative confidence interval, Az% 0,65
Critical values for convergence of resultsAz% < 4,8 < 4,8 Performed
Criterion statistical insignificance bias 5, % = | Z -100|< Az / < 0,265 Performed
Criterion practical significance of bias5, % = | Z -100|< 0,32^AAs < 1,536 Performed
The overall conclusion about the method Correct
Research robustness
To study this index was chosen such parameter as the stability of the sample solution in time (prepared by the methodology). As HFCs are not regulated through the period
Examining linearity, precision and accuracy of methods ranging from 50 to 150% of the nominal content loratadine, one could argue that the technique can withstand the requirements of HFCs, on a range of applications - for the quantitative determination of substances of medicines or dosage forms of 80 to 120% of the nominal content. So we developed a spectrophotometric method of quantitative determination of loratadine in tablets, conducted its validation. The basic validation parameters were specificity, linearity, range, accuracy (convergence), accuracy and robustness. Since fulfilled the eligibility criteria in respect of performance validation in accordance with the requirements of SPU, the method can be used for the determination of loratadine in tablets and can be included in the quality control methods for loratadine in tablets.
Conclusions:
1.developed spectrophotometric method of assay of loratadine in tablets. Validation of methods of quantitative determination of loratadine in tablets was conducted.
2.the validation parameters investigated methods of quantitative determination were specificity, linearity, range, accuracy (convergence), correctness, robustness.
3.since fulfilled the eligibility criteria in respect of performance validation in accordance with the requirements of HFCs, then this technique can be recommended to determine
of time necessary measure after preparation of the solution, so we chose a range of time - 2 hours. The results are shown in table 8, At< maxS = 1,02%, so the solution is stable for 2 hours (table 8).
the quantitative content APIs in tablets and included in the quality control methods for loratadine in tablets.
References
1)Korobko D.B., Polyauk O.B., Logoyda L.S et al. Development and validation of methods for identification of loratadine tablets. Actual questions of pharmaceutical and medical science and practice. (2014) 1:55-58.
2) State Pharmacopoeia of Ukraine: in 3 vol. / State Enterprise «Ukrainian Scientific Center pharmacopoeia quality medicines». - 2nd ed. - Kharkiv: State Enterprise «Ukrainian Scientific Center of Quality pharmacopoeia of medicines» (2014) 2:724.
3)State Pharmacopoeia of Ukraine / State Enterprise «Scientific and Expert Centre pharmacopoeia». - 1st ed. -Appendix 2 - Kharkiv: State Enterprise «Ukrainian Scientific Center of Quality pharmacopoeia of medicines» (2008):620.
4)Grizodub A.I. Reproducibility of pharmacopoeial methods of spectrophotometric quantitation of medicines in different laboratories. Farmakom. (2004) 2:20-34.
5) Grizodub A.I. Standard procedures validation techniques drug of quality control. Farmakom. (2006) 1-2:35-44.
6)Guidelines for the validation of methods of analysis of medicines / Under. Ed. N.V Yurgel, A.L Mladentsev. - Moscow (2007):57.
Table 8
Study of the stability of the test solution and reference solution
№ t, min Average RSDt, % At, % max5, %
0 20 40 60 80 100 120
1 0,572 0,569 0,568 0,568 0,568 0,569 0,569 0,569 0,224 0,31 1,02
2 0,569 0,569 0,569 0,569 0,569 0,569 0,569 0,569 0,044 0,06
